LOCUS       Exported                5667 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTJ1, complete sequence.
ACCESSION   JX559783
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  AUTHORS   Damron FH, McKenney ES, Schweizer HP, Goldberg JB.
  TITLE     Construction of a Broad-Host-Range Tn7-Based Vector for Single-Copy 
            PBAD-Controlled Gene Expression in Gram-Negative Bacteria
  JOURNAL   Appl. Environ. Microbiol. 79 (2), 718-721 (2013)
  PUBMED    23124231
REFERENCE   2  (bases 1 to 5667)
  AUTHORS   Damron FH, McKenney ES, Schweizer H, Goldberg JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-AUG-2012) Microbiology, Immunology, and Cancer 
            Biology, University of Virginia, PO Box 800734, Charlottesville, VA 
            22908, USA
REFERENCE   3  (bases 1 to 5667)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5667
                     /organism="Cloning vector pTJ1"
                     /lab_host="Escherichia coli strain TOP10"
                     /mol_type="other DNA"
                     /isolation_source="TOP10"
                     /db_xref="taxon:1229267"
     CDS             complement(7..885)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="AFR43359.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(7..885)
                     /gene="araC"
                     /label=araC
     promoter        912..1196
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      1157..1170
                     /regulatory_class="minus_35_signal"
                     /note="from PBAD"
     primer_bind     1171..1192
                     /label=pHERDSF primer
                     /note="pHERDSF primer"
     regulatory      1181..1188
                     /regulatory_class="minus_10_signal"
                     /note="from PBAD"
     RBS             1223..1245
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      1236..1241
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1255..1347
                     /note="NcoI-EcoR1-KpnI-SmaI-PstI-HindIII-NotI-KpnI multiple
                     cloning site"
     misc_feature    1257..1259
                     /label=start codon
                     /note="start codon"
     misc_feature    1271..1327
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     mobile_element  1368..1533
                     /mobile_element_type="transposon:Tn7L"
     oriT            1685..1953
     oriT            1692..1800
                     /note="incP origin of transfer"
     rep_origin      complement(2032..2620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2791..3651)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /protein_id="AFR43358.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2791..3651)
                     /gene="bla"
                     /label=bla
     promoter        complement(3652..3756)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  4234..4431
                     /mobile_element_type="transposon:Tn7R"
     primer_bind     4436..4452
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4480..4574
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      4677..4763
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     repeat_region   4814..4861
                     /rpt_family="FRT"
     protein_bind    4814..4861
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(4923..5159)
                     /codon_start=1
                     /gene="dhfRII"
                     /product="DhfRII"
                     /label=dhfRII
                     /protein_id="AFR43357.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     gene            complement(4923..5159)
                     /gene="dhfRII"
                     /label=dhfRII
     promoter        complement(5484..5512)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     repeat_region   5564..5611
                     /rpt_family="FRT"
     protein_bind    5564..5611
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."